通过Nutlin-3a激活P53揭示P53在hipsc心脏分化过程中ROS信号传导中的作用

Journal of stem cell reports Pub Date : 2021-01-01 Epub Date: 2021-03-27
Emma B Brandt, Xing Li, Timothy J Nelson
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引用次数: 0

摘要

癌细胞中转录因子P53的激活是一个很好的表征途径,而P53在发育过程中激活的影响在很大程度上仍未被探索。先前的研究表明,在小鼠发育的关键阶段,P53蛋白水平的增加会导致包括心脏在内的多个胚胎组织的缺陷。这些发现在几种不同的先天性心脏缺陷小鼠模型中得到了证实,但P53在人类心血管发育系统中的激活尚不清楚。利用人类诱导多能干细胞(hiPSCs),我们表征了心脏分化过程中P53的正常水平,并表明P53水平在hiPSCs中高,在心脏谱系承诺时降低。我们还观察到,在Nkx2-5 +心脏祖细胞阶段,P53的定位从iPS集落的主要细胞质转变为细胞核。在心脏发生的早期(中胚层到心脏中胚层)阶段,Mdm2抑制剂Nutlin-3a通过药物介导的P53蛋白水平升高导致心肌细胞凋亡增加和细胞周期停滞,从而导致心肌细胞大量损失。有趣的是,在心脏分化的后期(心脏祖细胞到跳动的心肌细胞),P53水平的增加并没有导致细胞凋亡。这些结果说明了在心脏发生过程中对P53水平升高的时间敏感性。我们对这些有或没有Nutlin-3a的细胞进行了RNA-Seq,以确定在分化的不同阶段由于P53增加而导致的转录差异。我们的RNA-Seq结果显示,在Nutlin-3a治疗后,Sestrins表达上调,这表明P53在心脏再生代谢中发挥了新的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Activation of P53 Via Nutlin-3a Reveals Role for P53 In ROS Signaling During Cardiac Differentiation of hiPSCs.

Activation of the transcription factor P53 within cancer cells is a well-characterized pathway, whereas the effects of P53 activation during development remain largely unexplored. Previous research has indicated that increased levels of P53 protein during key murine developmental stages cause defects in multiple embryonic tissues, including the heart. These findings were confirmed in several different mouse models of congenital heart defects, but P53 activation in a human system of cardiovascular development is not available. Utilizing human induced pluripotent stem cells (hiPSCs), we characterized the normal levels of P53 during cardiac differentiation and showed that levels of P53 are high in hiPSCs and decrease upon cardiac lineage commitment. We also observed P53 localization changed from mainly cytoplasmic in iPS colonies to the nucleus in the Nkx2-5 + cardiac progenitor stage. Pharmacological-mediated increase of P53 protein levels with the Mdm2 inhibitor Nutlin-3a during early (mesoderm to cardiac mesoderm) stages of cardiogenesis resulted in a sizeable loss of cardiomyocytes due to increased apoptosis and cell cycle arrest. Interestingly, increasing P53 levels did not result in apoptosis at later (cardiac progenitor to beating cardiomyocytes) stages of the cardiac differentiation. These results illustrate the temporal sensitivity to increased P53 levels during cardiogenesis. We conducted RNA-Seq on these cells with or without Nutlin-3a to ascertain transcriptional differences due to increased P53 at the different stages during the differentiation. Our results from the RNA-Seq revealed up-regulation of Sestrins after Nutlin-3a treatment suggesting a new role for P53 in the metabolism of cardiac regeneration.

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